使用敲除细胞株进行验证RabMAb

Anti-Integrin beta 1抗体[EP1041Y] (ab52971)

概述

  • 产品名称Anti-Integrin beta 1抗体[EP1041Y]
    参阅全部 Integrin beta 1 一抗
  • 描述
    兔单克隆抗体[EP1041Y] to Integrin beta 1
  • 经测试应用适用于: WB, IHC-Fr, IHC-Pmore details
    不适用于: Flow Cyt or ICC/IF
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Integrin beta 1 aa 650-750. The exact sequence is proprietary.
    Database link: P05556

  • 阳性对照
    • WB: HeLa, HT-1080, U937, U87-MG and U2OS whole cell lysates. IHC-P: Human transitional cell carcinoma of bladder and human breast cancer metastasis tissues.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

    Alternative versions available:

    Anti-Integrin beta 1 antibody (Alexa Fluor® 488) [EP1041Y] (ab190209)

    Anti-Integrin beta 1 antibody (Alexa Fluor® 647) [EP1041Y] (ab190595)

性能

应用

Our Abpromise guarantee covers the use of ab52971 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000 - 1/100000. Detects a band of approximately 140-150 kDa (predicted molecular weight: 88 kDa).

For unpurified, use 1/500.

IHC-Fr Use at an assay dependent concentration.
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Use of an HRP/AP polymerized secondary antibody is recommended.

For unpurified, use 1/250 - 1/500.

  • 应用说明Is unsuitable for Flow Cyt or ICC/IF.
  • 靶标

    • 功能Integrins alpha-1/beta-1, alpha-2/beta-1, alpha-10/beta-1 and alpha-11/beta-1 are receptors for collagen. Integrins alpha-1/beta-1 and alpha-2/beta-2 recognize the proline-hydroxylated sequence G-F-P-G-E-R in collagen. Integrins alpha-2/beta-1, alpha-3/beta-1, alpha-4/beta-1, alpha-5/beta-1, alpha-8/beta-1, alpha-10/beta-1, alpha-11/beta-1 and alpha-V/beta-1 are receptors for fibronectin. Alpha-4/beta-1 recognizes one or more domains within the alternatively spliced CS-1 and CS-5 regions of fibronectin. Integrin alpha-5/beta-1 is a receptor for fibrinogen. Integrin alpha-1/beta-1, alpha-2/beta-1, alpha-6/beta-1 and alpha-7/beta-1 are receptors for lamimin. Integrin alpha-4/beta-1 is a receptor for VCAM1. It recognizes the sequence Q-I-D-S in VCAM1. Integrin alpha-9/beta-1 is a receptor for VCAM1, cytotactin and osteopontin. It recognizes the sequence A-E-I-D-G-I-E-L in cytotactin. Integrin alpha-3/beta-1 is a receptor for epiligrin, thrombospondin and CSPG4. Alpha-3/beta-1 may mediate with LGALS3 the stimulation by CSPG4 of endothelial cells migration. Integrin alpha-V/beta-1 is a receptor for vitronectin. Beta-1 integrins recognize the sequence R-G-D in a wide array of ligands. Isoform 2 interferes with isoform 1 resulting in a dominant negative effect on cell adhesion and migration (in vitro). When associated with alpha-7/beta-1 integrin, regulates cell adhesion and laminin matrix deposition. Involved in promoting endothelial cell motility and angiogenesis. Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process and the formation of mineralized bone nodules. May be involved in up-regulation of the activity of kinases such as PKC via binding to KRT1. Together with KRT1 and RACK1, serves as a platform for SRC activation or inactivation. Plays a mechanistic adhesive role during telophase, required for the successful completion of cytokinesis. Integrin alpha-3/beta-1 provides a docking site for FAP (seprase) at invadopodia plasma membranes in a collagen-dependent manner and hence may participate in the adhesion, formation of invadopodia and matrix degradation processes, promoting cell invasion. ITGA4:ITGB1 binds to fractalkine (CX3CL1) and may act as its coreceptor in CX3CR1-dependent fractalkine signaling (PubMed:23125415, PubMed:24789099). ITGA4:ITGB1 and ITGA5:ITGB1 bind to PLA2G2A via a site (site 2) which is distinct from the classical ligand-binding site (site 1) and this induces integrin conformational changes and enhanced ligand binding to site 1 (PubMed:18635536, PubMed:25398877). ITGA5:ITGB1 acts as a receptor for fibrillin-1 (FBN1) and mediates R-G-D-dependent cell adhesion to FBN1 (PubMed:12807887, PubMed:17158881).
      Isoform 5: Isoform 5 displaces isoform 1 in striated muscles.
      (Microbial infection) Integrin ITGA2:ITGB1 acts as a receptor for human echoviruses 1 and 8 (PubMed:8411387). Acts as a receptor for cytomegalovirus/HHV-5 (PubMed:20660204). Acts as a receptor for Epstein-Barr virus/HHV-4 (PubMed:17945327). Integrin ITGA5:ITGB1 acts as a receptor for human parvovirus B19 (PubMed:12907437). Integrin ITGA2:ITGB1 acts as a receptor for human rotavirus (PubMed:12941907). Acts as a receptor for mammalian reovirus (PubMed:16501085). In case of HIV-1 infection, integrin ITGA5:ITGB1 binding to extracellular viral Tat protein seems to enhance angiogenesis in Kaposi's sarcoma lesions (PubMed:10397733).
    • 组织特异性Isoform 1 is widely expressed, other isoforms are generally coexpressed with a more restricted distribution. Isoform 2 is expressed in skin, liver, skeletal muscle, cardiac muscle, placenta, umbilical vein endothelial cells, neuroblastoma cells, lymphoma cells, hepatoma cells and astrocytoma cells. Isoform 3 and isoform 4 are expressed in muscle, kidney, liver, placenta, cervical epithelium, umbilical vein endothelial cells, fibroblast cells, embryonal kidney cells, platelets and several blood cell lines. Isoform 4, rather than isoform 3, is selectively expressed in peripheral T-cells. Isoform 3 is expressed in non-proliferating and differentiated prostate gland epithelial cells and in platelets, on the surface of erythroleukemia cells and in various hematopoietic cell lines. Isoform 5 is expressed specifically in striated muscle (skeletal and cardiac muscle).
    • 序列相似性Belongs to the integrin beta chain family.
      Contains 1 VWFA domain.
    • 翻译后修饰The cysteine residues are involved in intrachain disulfide bonds.
    • 细胞定位Cell membrane, sarcolemma. Cell junction. In cardiac muscle, isoform 5 is found in costameres and intercalated disks and Cell membrane. Cell projection, invadopodium membrane. Cell projection, ruffle membrane. Recycling endosome. Melanosome. Cleavage furrow. Cell projection, lamellipodium. Cell projection, ruffle. Cell junction, focal adhesion. Cell surface. Isoform 2 does not localize to focal adhesions. Highly enriched in stage I melanosomes. Located on plasma membrane of neuroblastoma NMB7 cells. In a lung cancer cell line, in prometaphase and metaphase, localizes diffusely at the membrane and in a few intracellular vesicles. In early telophase, detected mainly on the matrix-facing side of the cells. By mid-telophase, concentrated to the ingressing cleavage furrow, mainly to the basal side of the furrow. In late telophase, concentrated to the extending protrusions formed at the opposite ends of the spreading daughter cells, in vesicles at the base of the lamellipodia formed by the separating daughter cells. Colocalizes with ITGB1BP1 and metastatic suppressor protein NME2 at the edge or peripheral ruffles and lamellipodia during the early stages of cell spreading on fibronectin or collagen. Translocates from peripheral focal adhesions sites to fibrillar adhesions in a ITGB1BP1-dependent manner. Enriched preferentially at invadopodia, cell membrane protrusions that correspond to sites of cell invasion, in a collagen-dependent manner. Localized at plasma and ruffle membranes in a collagen-independent manner.
    • Information by UniProt
    • 数据库链接
    • 别名
      • beta1 integrin antibody
      • CD29 antibody
      • Fibronectin receptor subunit beta antibody
      • FNRB antibody
      • Glycoprotein IIa antibody
      • GP IIa antibody
      • GPIIA antibody
      • Integrin beta-1 antibody
      • integrin VLA-4 beta subunit antibody
      • Integrin, beta 1 (fibronectin receptor, beta polypeptide, antigen CD29 includes MDF2, MSK12) antibody
      • ITB1_HUMAN antibody
      • ITGB1 antibody
      • MDF2 antibody
      • MSK12 antibody
      • OTTHUMP00000019420 antibody
      • Very late activation protein, beta polypeptide antibody
      • VLA BETA antibody
      • VLA-4 subunit beta antibody
      • VLA-BETA antibody
      • VLAB antibody
      • VLAbeta antibody
      see all

    Anti-Integrin beta 1 antibody [EP1041Y] 图像



    • Predicted band size : 88 kDa

      Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
      Lanes 2, 6 and 10: Integrin beta 1 knockout HAP1 cell lysate (20 µg)
      Lanes 3, 7 and 11: U87-MG cell lysate (20 µg)
      Lanes 4, 8 and 12: A431 cell lysate (20 µg)
      Lanes 1, 2, 3 and 4: Green signal from target – ab52971 observed at 140 kDa
      Lanes 5, 6, 7 and 8: Red signal from loading control – ab8245 observed at 37 kDa
      Lanes 9, 10, 11 and 12: Merged (red and green) signal

      ab52971 was shown to specifically react with Integrin beta 1 when Integrin beta 1 knockout samples were used. Wild-type and Integrin beta 1 knockout samples were subjected to SDS-PAGE. ab52971 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • ab52971 at 1/500 staining Integrin beta 1 antibody in human transitional cell carcinoma of bladder by immunohistochemistry (FFPE).

      Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling Integrin beta 1 with ab52971 at 1/500 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051, 1/500). Counter stained with hematoxylin.

    • All lanes : Anti-Integrin beta 1 antibody [EP1041Y] (ab52971) at 1/10000 dilution (purified)

      Lane 1 : U87-MG cell lysate
      Lane 2 : HT-1080 cell lysate
      Lane 3 : U937 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 88 kDa
      Observed band size : 140 kDa (why is the actual band size different from the predicted?)

      5% NFDM/TBST dilution buffer

    • All lanes : Anti-Integrin beta 1 antibody [EP1041Y] (ab52971) at 20 µg (unpurified)

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : HT 1080 (Human fibrosarcoma) Whole Cell Lysate
      Lane 3 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution

      Predicted band size : 88 kDa
      Observed band size : 120 + 140 kDa (why is the actual band size different from the predicted?)

      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab52971 overnight at 4°C. Antibody binding was detected using ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

       

      Secondary antibody - anti-rabbit Alexa Fluor 790

    • Unpurified ab52971 staining human breast cancer metastasis tissue sections by IHC-P. Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking with a commercial blocking reagent and incubation with the antibody (diluted 1/100) for 18 hours at 4°C.  A HRP-conjugated goat anti-rabbit was used as the secondary antibody.  This image shows a cancer metastasis at 40x with beta1 staining (in red) in both blood vessels and tumour cells. Blue is Hoechst for nuclei.  The antibody detection was enhanced using a commercial Cy3 tyramide signal amplification kit.

      See Abreview

    Anti-Integrin beta 1 antibody [EP1041Y] (ab52971)参考文献

    This product has been referenced in:
    • Hatanaka K  et al. Molecular mechanisms of the angiogenic effects of low-energy shock wave therapy: roles of mechanotransduction. Am J Physiol Cell Physiol 311:C378-85 (2016). Read more (PubMed: 27413171) »
    • Zhao W  et al. Effect of TGF-ß1 on the Migration and Recruitment of Mesenchymal Stem Cells after Vascular Balloon Injury: Involvement of Matrix Metalloproteinase-14. Sci Rep 6:21176 (2016). IHC-P ; Rat . Read more (PubMed: 26880204) »

    See all 31 Publications for this product

    Product Wall

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunohistochemistry (Frozen sections)
    Sample Medaka fish Tissue sections (Basal side of epithelia)
    Specification Basal side of epithelia
    Fixative DMSO+Methanol
    Permeabilization Yes - acetone
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Username

    Dr. Makoto Furutani-Seiki

    Verified customer

    提交于 Nov 15 2011

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Human Tissue sections (Primary humain airway epithelial cells)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate biffer 10mM
    Permeabilization No
    Specification Primary humain airway epithelial cells
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Fixative Formaldehyde
    Username

    Dr. Aaron Gardner

    Verified customer

    提交于 Nov 30 2016

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (4-15%)
    Sample Human Cell lysate - whole cell (non small cell lung cancer)
    Specification non small cell lung cancer
    Treatment siRNA treated
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 Oct 07 2014

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step FBS / BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris pH 9
    Sample Human Tissue sections (Infantile fibromatosis)
    Specification Infantile fibromatosis
    Permeabilization No
    Fixative Formaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Jul 21 2014

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step 1% serum + 0.1% BSA in PBS as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 16°C
    Sample Mouse Cell (mouse embryonic stem cells)
    Specification mouse embryonic stem cells
    Permeabilization Yes - 0.1% triton X
    Fixative methanol:acetone
    Username

    Abcam user community

    Verified customer

    提交于 Apr 30 2014

    Application Western blot
    Loading amount 1e+006 cells
    Gel Running Conditions Reduced Denaturing (10% gel)
    Sample Mouse Cell lysate - whole cell (mouse embryonic stem cells)
    Specification mouse embryonic stem cells
    Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
    Username

    Abcam user community

    Verified customer

    提交于 Nov 28 2013

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
    Sample Human Cell (HeLa cell)
    Specification HeLa cell
    Permeabilization Yes - 0.5% TritonX/PBS
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Nov 19 2013

    Please forward the following publication to customer, there are many markers listed and we have antibodies against these. Customer might need to make decision based on their requirements.

    http://journals.tums.ac.ir/upload_files/pdf/_/20047.pdf...

    Read More
    Application Immunocytochemistry/ Immunofluorescence
    Sample Mouse Cell (Primary Cortical Neuron Culture)
    Specification Primary Cortical Neuron Culture
    Fixative Paraformaldehyde
    Permeabilization Yes - Triton 0.25%
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
    Username

    Dr. Sofia Santos

    Verified customer

    提交于 Apr 24 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step 1% BSA 1% FBS as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris EDTA pH 9
    Sample Mouse Tissue sections (mouse vein)
    Specification mouse vein
    Permeabilization Yes - Tween-20
    Fixative Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    提交于 Dec 12 2012

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